So Nishimoto, Teisuke Takita, Kazumichi M Nishida, Masaaki Ito, Toshiyuki Kimura, Yasuhisa Kimura, Noriyuki Kioka, Kiyoshi Yasukawa
{"title":"重组人α-葡萄糖苷酶在HEK293细胞中的表达、纯化和表征。","authors":"So Nishimoto, Teisuke Takita, Kazumichi M Nishida, Masaaki Ito, Toshiyuki Kimura, Yasuhisa Kimura, Noriyuki Kioka, Kiyoshi Yasukawa","doi":"10.1093/bbb/zbaf055","DOIUrl":null,"url":null,"abstract":"<p><p>In mammals, intestinal α-glucosidase exists within maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI) complexes. Here, we purified recombinant human MGAM and SI from human embryonic kidney 293 (HEK293) cells using Flag tag-based affinity chromatography. Steady-state kinetic analysis at pH 6.0 and 37°C showed that the maltase activities of MGAM and SI were higher than their sucrase and isomaltase activities.</p>","PeriodicalId":9175,"journal":{"name":"Bioscience, Biotechnology, and Biochemistry","volume":" ","pages":""},"PeriodicalIF":1.4000,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Expression in HEK293 cells, purification, and characterization of recombinant human α-glucosidases.\",\"authors\":\"So Nishimoto, Teisuke Takita, Kazumichi M Nishida, Masaaki Ito, Toshiyuki Kimura, Yasuhisa Kimura, Noriyuki Kioka, Kiyoshi Yasukawa\",\"doi\":\"10.1093/bbb/zbaf055\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>In mammals, intestinal α-glucosidase exists within maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI) complexes. Here, we purified recombinant human MGAM and SI from human embryonic kidney 293 (HEK293) cells using Flag tag-based affinity chromatography. Steady-state kinetic analysis at pH 6.0 and 37°C showed that the maltase activities of MGAM and SI were higher than their sucrase and isomaltase activities.</p>\",\"PeriodicalId\":9175,\"journal\":{\"name\":\"Bioscience, Biotechnology, and Biochemistry\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":1.4000,\"publicationDate\":\"2025-04-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Bioscience, Biotechnology, and Biochemistry\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1093/bbb/zbaf055\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bioscience, Biotechnology, and Biochemistry","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1093/bbb/zbaf055","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Expression in HEK293 cells, purification, and characterization of recombinant human α-glucosidases.
In mammals, intestinal α-glucosidase exists within maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI) complexes. Here, we purified recombinant human MGAM and SI from human embryonic kidney 293 (HEK293) cells using Flag tag-based affinity chromatography. Steady-state kinetic analysis at pH 6.0 and 37°C showed that the maltase activities of MGAM and SI were higher than their sucrase and isomaltase activities.
期刊介绍:
Bioscience, Biotechnology, and Biochemistry publishes high-quality papers providing chemical and biological analyses of vital phenomena exhibited by animals, plants, and microorganisms, the chemical structures and functions of their products, and related matters. The Journal plays a major role in communicating to a global audience outstanding basic and applied research in all fields subsumed by the Japan Society for Bioscience, Biotechnology, and Agrochemistry (JSBBA).